Summary
The effects of poly-l-ornithine on the surface membrane of isolated tobacco protoplasts have been examined in the electron microscope using a colloidal metal oxide and a spherical virus as marker substances. No evidence was found to suggest that isolated protoplasts take up either of these markers by a pinocytotic process. Poly-l-ornithine increased the degree of damage observed in fixed preparations, and specifically caused lesions of the plasmalemma which were favoured sites for the binding of both external marker substances. It is suggested that the function of poly-l-ornithine and other treatments used to obtain virus infection of protoplasts is to stress the cell membrane to allow a non-physiological entry of high molecular weight materials. Pinocytosis appears not to occur nor to be necessary for uptake of these materials under conditions of membrane stress.
Similar content being viewed by others
References
Aoki, S., Takebe, I.: Infection of tobacco mesophyll protoplasts by tobaceo mosaic virus ribonucleic acid. Virology 39, 439–448 (1969).
Bancroft, J. B., Hills, G. J., Markham, R.: A study of the self-assembly process in a small spherical virus. Formation of organised structures from protein subunits in vitro. Virology 31, 354–379 (1967).
Bessis, M.: Studies on cell agony and death; an attempt at classification. Ciba Foundation Symposium, “Cellular injury”, ed. A. V. S. de Reuck and J. Knight. London: Churchill 1964.
Bradfute, O. E., Chapman-Andersen, C., Jensen, W. A.: Concerning morphological evidence for pinocytosis in higher plants. Exp. Cell Res. 36, 207–210 (1964).
Burgess, J., Watts, J. W., Fleming, E. N., King, J. M.: Plasmalemma fine structure in isolated tobacco mesophyll protoplasts. Planta (Berl.), 110, 291–301 (1973).
Cocking, E. C.: An electron microscopic study of the initial stages of infection of isolated tomato fruit protoplasts by TMV. Planta (Berl.) 68, 206–214 (1966).
Cocking, E. C., Pojnar, E.: An electron microscopic study of the infection of isolated tomato fruit protoplasts by TMV. J. gen. Virol. 4, 305–312 (1969).
Coutts, R. H. A., Cocking, E. C., Kassanis, B.: Infection of tobacco mesophyll protoplasts with TMV. J. gen. Virol. 17, 289–294 (1972).
Drew, M. C., Seear, J., McLaren, A. D.: Entry of basic macromolecules into barley roots. Amer. J. Bot. 57, 837–843 (1970).
Fawcett, D. W.: Surface specialisations of absorbing cells. J. Histochem. Cytochem. 13, 75–91 (1965).
Hibi, T., Yora, K.: Electron microscopy of tobacco mosaic virus infection in tobacco mesophyll protoplasts. Ann. Phytopath. Soc. Japan 38, 350–356 (1972).
Kent, S. P.: Intracellular plasma protein: a manifestation of cell injury in myocardial ischaemia. Nature (Lond.) 210, 1279–1281 (1966).
Lagunoff, D., Curran, D. E.: Role of bristle-coated membrane in the uptake of ferritin by rat macrophages. Exp. Cell Res. 75, 337–346 (1972).
Mahlberg, P., Olson, K., Walkinshaw, C.: Origin and development of plasmalemma derived invaginations in Vinca rosea L. Amer. J. Bot. 58, 409–416 (1971).
Mayo, M. A., Cocking, E. C.: Pinocytotic uptake of polystyrene latex particles by isolated tomato fruit protoplasts. Protoplasma (Wien) 68, 223–230 (1969).
Otsuki, Y., Takebe, I., Honda, Y., Matsui, C.: Ultrastructure of infection of tobacco mesophyll protoplasts by tobacco mosaic virus. Virology 49, 188–194 (1972).
Oudea, P. R.: Anoxic changes of liver cells. Lab. Invest. 12, 386–394 (1963).
Pojnar, E., Cocking, E. C.: The uptake of viruses, cell wall regeneration and virus multiplication in isolated plant protoplasts. Proceedings of the 2nd Int. Symp. on Yeast Protoplasts, ed. Necas, O., and A. Svoboda (1970).
Prat, R.: Contribution a l'étude des protoplastes végétaux. 1. Effet du traitement d'isolement sur la structure cellulaire. J. Microscopie 14, 85–114 (1972).
Ryser, H.: Studies on protein uptake by isolated tumour cells. III. Apparent stimulation due to pH, hypertonicity, polycations and dehydration, and their relation to the enhanced penetration of infections nucleic acids. J. Cell Biol. 32, 737–750 (1967).
Ryser, H., Caulfield, J. B., Aub, J. C.: Studies on protein uptake by isolated tumour cells. 1. Electron-microscopic evidence of ferritin uptake by Ehrlich ascites tumour cells. J. Cell Biol. 14, 255–268 (1962).
Ryser, H., Hancock, R.: Histones and basic polyamino acids stimulate the uptake of albumin by tumour cells in culture. Science 150, 501–503 (1965).
Seear, J., Bradfute, O. E., McLaren, A. D.: Uptake of proteins by plant roots. Phys. Plant. 21, 979–989 (1968).
Takebe, I., Otsuki, Y.: Infection of tobacco mesophyll protoplasts by tobacco mosaic virus. Proc. nat. Acad. Sci. (Wash.) 64, 843–848 (1969).
Takebe, I., Otsuki, Y., Aoki, S.: Isolation of tobacco mesophyll cells in intact and active state. Plant & Cell Physiol. 9, 115–124 (1968).
Wheeler, H., Baker, B. L., Hanchey, P.: Pinocytosis in root cap cells exposed to uranyl salts. Amer. J. Bot. 59, 858–868 (1972).
Wheeler, H., Hanchey, P.: Pinocytosis and membrane dilation in uranyl treated plant roots. Science 171, 68–71 (1971).
Withers, L. A., Cocking, E. C.: Fine structural studies on spontaneous and induced fusion of higher plant protoplasts. J. Cell Sci. 11, 59–75 (1972).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Burgess, J., Motoyoshi, F. & Fleming, E.N. Effect of poly-l-ornithine on isolated tobacco mesophyll protoplasts: Evidence against stimulated pinocytosis. Planta 111, 199–208 (1973). https://doi.org/10.1007/BF00385104
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00385104